1. Field of the Invention
The present invention relates to a hybridoma cell line, particularly to a hybridoma cell line which produces an antibody which specifically binds to a neopeptide of type II collagen and a method for detecting osteoarthritis by identifying a presence of type II collagen in a urine sample via the said antibody.
2. Description of the Related Art
Soft tissue of joint refers to tissues that connect, support, and surround bones, to cushion the friction between bones. There is a dynamic equilibrium between the synthesis and degradation of the molecules of the soft tissue of joint, for maintaining the said functions of the soft tissue of joint.
Degenerative arthritis, also known as osteoarthritis (OA), is a group of mechanical abnormalities involving degradation and deform of articular cartilage, and joint pain, tenderness or stiffness, caused by disequilibrium between the synthesis and degradation of the soft tissue of joint.
Conventional detections of osteoarthritis generally bases on identifying degraded fragments of soft tissues, such as collagen and proteoglycans in body fluids. The degraded fragments of soft tissues are present only when joints suffer from excessive pressures or friction. Due to the specificity of the degraded fragments of soft tissues, it is capable of being a biomarker of the detections of osteoarthritis. The conventional degraded fragments of soft tissues comprise CTXII, HELIX-II, C2C, coll2-1 and collagenase recognized site, including C2C1 and TII NE, and which have been widely used in screening people that is apt to have osteoarthritis.
A conventional method for detecting osteoarthritis comprises steps of identifying a presence of C2C fragment in blood samples or other body fluids, and detecting the blood samples or other body fluids with an antibody against C2C fragment. However, the titer and sensitivity of the antibody against C2C fragment is poor, only can being applied to samples that is collected via invasive procedures, blood and synovial fluid for example, since the C2C fragment has more than 20 amino acids, being easy to activate immune response in model animal and to prepare antibodies against thereof, but being inconvenient in practical use at home.
For improving the above conventional method, another conventional method for detecting osteoarthritis is disclosed in U.S. Pat. No. 6,642,007, entitled of “ASSAYS FOR MEASUREMENT OF TYPE II COLLAGEN FRAGMENTS IN URINE.” The said conventional method for detecting osteoarthritis is carried by identifying a presence of type II collagen in urine sample with a complex of antibodies, comprising a capture antibody and a detection antibody against that against different epitopes on type II collagen neoepitope (TII NE), with the capture and detection antibodies both binding to target sequences, so that the sensitivity of the capture antibody can be promoted.
In specific, the said conventional method for detecting osteoarthritis is performed via sandwich enzyme linked immunosorbent assay, for improving the poor sensitivity of single antibody. However, the said conventional method for detecting osteoarthritis needs more resources, costs and times to develop various antibodies that against more than one epitope, and thus that, the diagnosis of osteoarthritis can not be achieved through an easy and low-cost process.
Hence there is a need of providing a user-friendly diagnostic method and apparatus for detecting osteoarthritis, which can provide an antibody that specifically binds to an epitope only comprising 10-15 amino acids from type II collagen, and does not bind to type I collagen or type III collagen.